Project Summary Ethanol-induced changes in the brain transcriptome underlie the development and persistence of alcohol use disorder (AUD). INIA-N investigators and others have discovered that ethanol induces specific and dramatic alterations in a highly restricted group of noncoding RNAs (ncRNAs) that includes long ncRNAs (lncRNAs). We posit that these ethanol responsive lncRNAs coordinate AUD brain transcriptomes. Specific Aim 1 will test the hypothesis that individual lncRNAs are key regulators of ethanol drinking. To test this hypothesis, genetically engineered mice with altered expression of lncRNAs will be created and analyzed. Those mutant lines with altered drinking behavior will be scrutinized for mechanistic insight by multiple INIA-N investigators. A barrier to the study of ncRNA function in brain is the dearth of efficient methods of noninvasively delivering ncRNAs and/or ncRNA antagonists to large portions of the brain. Here we posit that intransally administered exosomes (endogenously produced, liposome like nanoparticles) can be harnessed to deliver ncRNA anatagonists or mimics throughout the brain. This approach can also be used to deliver drugs preferentially to brain. Such an approach would target drugs selectively to the desired site of action (brain) while avoiding peripheral toxicities that limit therapeutic efficacy. Because this approach has the dual benefits of target specificity and noninvasiveness, it has tremendous translational potential. Specific Aim 2 will test the hypothesis that exosomes can be harnessed as effective ncRNA/drug delivery vehicles to modulate ethanol drinking. Genetically engineered rodents permit investigation of the involvement of putative ethanol targets in the context of whole animal behavioral responses. Because hypotheses concerning putative ethanol targets must ultimately explain ethanol-induced behavioral phenotypes, whole-animal experiments represent the most rigorous test of relevance. To this end, Specific Aim 3 will create designer mice for both INIA-N and INIA-Stress investigators. Genetically engineered animals will be produced using state of the art CRISPR/Cas9 gene editing technology. This collaborative Specific Aim is the continuation of the INIA-West Genetically Engineered Rodents Core that was funded during the previous project period.